The overall goal of the INIA-stress consortium is to integrate efforts to define the allostatic state of the cortical-limbic-HPA axis produced by excessive alcohol consumption as a platform for studying the relationship between stress, the subjective state of anxiety, and excessive drinking. The objective of the Gene-Targeted Mouse Core is to create novel animal models that allow spatial and/or temporal disruption of specific brain targets. These targets are receptors or their ligands that play key roles in modulating biological attributes such as anxiety, reward, and emotional behavior. Circuitries underlying these attributes are all affected by alcohol abuse. The use of targeted gene disruption will allow the dissection of individual pathways in relationship to alcohol consumption and alcoholism. For this renewal, the Core has been tasked to create seven new mouse models while taking advantage of available gene-targeted embryonic stem cell lines generated by the European Knockout Consortium and the Knockout Mouse Project (KOMP). When available, the ES cell lines will be obtained and injected into C57 albino blastocysts. Homozygous floxed mice will be mated to mice carrying inducible, forebrain-specific recombinase transgenes. For targets with unavailable ES cell lines, the Core will engineer a targeting construct using BAC recombineering techniques and target ES cells for recombination. Once completed, inducible knockout mice will be shipped to the Neuroanatomy Phenotyping Core for anatomical analyses and a larger number of breeders will be shipped to the Mouse Chronic Intermittent Ethanol (CIE) Core for alcohol phenotyping. The Core will also breed, maintain, distribute to INIA-stress investigators, and cryopreserve all mouse lines. Finally, the Core will provide genotyping services to INA-stress investigators. PUBLIC HEALTH RELEVANCE: The Gene-Targeted Mouse Core creates novel inducible-knockout mouse models for INIA-stress investigators and ultimately for the larger research community. The targets, selected by the INIA-stress consortium are receptors or their ligands that play key roles in modulating biological attributes such as anxiety, reward, and emotional behavior. The use of targeted gene disruption allows the dissection of individual pathways in relationship to alcohol consumption and alcoholism.